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表面等离子体共振免疫分析——综述

Surface plasmon resonance immuno assays - A perspective.

作者信息

Gutiérrez-Gallego R, Bosch J, Such-Sanmartín G, Segura J

机构信息

Bioanalysis and Analytical Services Research Group, Neuropsychopharmacology Program, Municipal Institute of Medical Research (IMIM-Hospital del Mar), PRBB, Barcelona, Spain.

出版信息

Growth Horm IGF Res. 2009 Aug;19(4):388-98. doi: 10.1016/j.ghir.2009.04.022. Epub 2009 May 26.

Abstract

Human growth hormone (GH) represents an extremely challenging task from an anti-doping viewpoint. GH is an endogenously produced substance, present at very low levels in circulation (for the most abundant 22kDa isoform approximately 50pM in plasma and 100fM in urine) either as monomer or homo- and heterodimers, comprises a family of distinct isoforms, and obeys a pulsatile secretion routine that is affected by many different internal and external factors. Upon administration of the recombinant, single-isoform pharmaceutical, the feedback mechanism reduces the endogenous heterogeneity resulting in altered ratios between the different GH isoforms. Thus, measuring the isoform ratios through immuno assays appears the approach of choice. Conventional assays do not provide information on isoform-specific association and dissociation events of the individual primary antibody-isoform or isoform-secondary antibody interactions. This particular information can be obtained using the technology of surface plasmon resonance (SPR) which enables monitoring of biomolecular interactions in a dynamic and label-free setting. In this paper the different aspects of SPR are described, how the technology may be beneficial for understanding today's anti-GH immunoassays, and whether the approach could be employed for measuring GH in the near future.

摘要

从反兴奋剂的角度来看,人类生长激素(GH)是一项极具挑战性的任务。生长激素是一种内源性产生的物质,以单体或同二聚体和异二聚体的形式存在于循环系统中的水平非常低(对于最丰富的22kDa异构体,血浆中约为50pM,尿液中约为100fM),它由一系列不同的异构体组成,并遵循受许多不同内部和外部因素影响的脉冲式分泌规律。在使用重组单异构体药物后,反馈机制会降低内源性异质性,导致不同生长激素异构体之间的比例发生变化。因此,通过免疫测定法测量异构体比例似乎是首选方法。传统测定法无法提供关于单个一抗-异构体或异构体-二抗相互作用的异构体特异性结合和解离事件的信息。使用表面等离子体共振(SPR)技术可以获得这些特定信息,该技术能够在动态且无标记的环境中监测生物分子相互作用。本文描述了表面等离子体共振的不同方面,该技术如何有助于理解当今的抗生长激素免疫测定法,以及该方法在不久的将来是否可用于测量生长激素。

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